• KOREAN JOURNAL OF CROP SCIENCE
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• v.27 no.2
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• pp.147-154
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• 1982

This study was conducted to investigate an effective method of protoplast isolation, the plating efficiency for cell division, and fusion of plant protoplasts by polyethylene glycol for somatic hybridzation. The effectiveness of protoplast isolation was different with the various enzyme concentrations, but, in the protoplast isolation from tobacco mesophyll, the enzyme solution with 0.5% macerozyme and 2.0% cellulase was very effective. The protoplast isolation from callus cultured in vitro for a long period was not obtained in any of the enzyme solution used. Protoplasts divided actively at cell densities above $10^44/ml and at $25^{\circ}C$ under 12hr illumination by inflorecient light (l50 Lux), regardless of presence of agar. The highest frequency of protoplast fusion was obtained after treatment with a solution of 0.33 M polyethylene glycol 1500.

• Applied Microscopy
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• v.38 no.4
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• pp.307-314
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• 2008

Hydrophytes such as Spirodela polyrhiza form dormant turions to withstand cold winters. The turion is an anatomically distinct structure from which a vegetative frond arises later during germination. The turions sink to the bottom of the pond when temperatures drop and remain there throughout the winter. In the spring, they float to the surface and germinate into a new frond from the turion primordium. Unlike fronds, turions are known to possess small aerenchyma, starch grains, and relatively dense cytoplasm. These features allow the turions to survive the cold winter season at the bottom of the pond. Spirodela polyrhiza has been investigated previously to a great extent, especially in its physiological, biochemical and ecological attributes. However, a little is known about the structural features of the frond and turion during turion development. Thus, the aim of the present study was to reveal the structural characteristics of the frond and turion with regard to tissue differentiation, aerenchyma development, starch distribution, and ultrastructure, with the use of electron microscopy. A moderate degree of mesophyll tissue differentiation was found in the frond, whereas the turion did not exhibit such differentiation. Within the frond tissue, approximately $37{\sim}45%$ of the cellular volume was occupied by a large aerenchyma, but only $9{\sim}15%$ was taken up by the aerenchyma in the turion. The turion cells, especially those of the turion primordium, were derived from frond cells, and contained cytoplasm. Their cytoplasm was densely packed with plastids, mitochondria, endoplasmic reticulum, Golgi bodies, and microtubules. Plasmodesmata were also well developed within these cells. The most striking feature observed was the distribution of starch grains within the plastids of turion cells. Before the turion sank to the bottom of the pond, a considerable amount of starch accumulated in the plastid stroma. The starch grains dissolved when temperatures rose in the spring, and this promptly provided the nutrients which the primordium needed for turion germination. The turion therefore, was an appropriate dormant structure for free-floating, reduced hydrophytes like Spirodela polyhriza due to its small aerenchyma and large starch grains that aided in the purpose of sinking below the surface of the water to survive cold winters. The new fronds that arose from such turions grew rapidly in the spring, beginning the new life cycle.

• Journal of Environmental Science International
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• v.17 no.2
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• pp.163-170
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• 2008

Protoplasts of Panax ginseng C. A. Meyer and Aralia continentalis K. (Araliaceae) were isolated from callus cells and mesophyll cells, respectively. The maximum yield of protoplasts isolated from callus cells of P. ginseng were obtained by incubation for 3 hrs in the enzyme mixture of 0.5% macerozyme, 1.5% cellulase, and 0.5 M mannitol as an osmoticum. In the case of mesophyll cells of A. continentalis, the highest yield of protoplasts were obtained by incubation for 5 hrs in the enzyme mixture of 1% macerozyme, 2% cellulase, and 0.6 M mannitol. A polyethylene glycol (PEG) treatment induced an intergeneric fusion of the protoplasts. The fusion products, that is, heterokaryocytes were obtained by treatment of 50% PEG containing 0.05 M Ca salts.

• Korean Journal Plant Pathology
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• v.12 no.3
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• pp.368-373
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• 1996

배나무잎 검은점병에 이병된 신고와 지표식물 PS-95의 잎을 전자현미경으로 세포내 미세구조를 검경한 결과 굴곡성 사상형 유사바이러스 입자가 집단으로 존재하고 있는 것을 확인하였다. 엽육유세포질에 있는 유사바이러스 입자들의 직경은 12 nm였으나 입자들의 길이는 측정하지 못하였다. 섬유사를 함유하고 있는 소포는 일반적으로 ssRNA genome을 갖는 식물바이러스에 의해 이병된 세포에서 생성된다. 본 연구에서 이 소포들은 tonoplast에 형성되었다. 배나무잎 검정점병의 이병잎을 초본 지표식물에 즙액접종하였으나 어떠한 병징도 나타나지 않았다. 또한 접목접종 전염에 의하여 전염되어 전형적인 검은점이 발병하였다. 발병된 잎에는 유사바이러스 입자가 존재하고 있었다. 이상의 결과 병징, 섬유사를 함유한 소포의 존재, 그리고 접목전염을 기초로하여 볼 때 배나무 검은점병을 일으키는 유사바이러스 입자는 closteroviruses의 하나로 생각된다.

• Journal of The Korean Society of Grassland and Forage Science
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• v.9 no.3
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• pp.119-123
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• 1989

레드 클로버의 葉肉細胞로 부터 原形質體의 裸出에 影響을 미치는 要因들을 調査하여 다음과 같은 結果를 얻었다. 原形質體의 裸出에 使用된 酵素는 Cellulase, Macerozyme, Driselase 및 Pectolyase였으며 그 中 1%의 Cellulase ONOZUKA R-10과 0.5%의 Macerozyme R-10의 組合에서 가장 좋은 結果를 얻었다. 裸出된 原形質體의 適正 膨壓을 維持하기 爲하여 Mannitol의 濃度를 0.3~O.8M의 범위에서 6단계로 조정하였을 때, 0.7M에서 가장 높은 收率을 얻을 수 있었다. 原形質體 裸出을 爲한 酵素溶液의 pH는 5.6에서 가장 좋은 結果를 나타내었으며, 材料로 쓰인 레드 클로버 일을 4가지 크기로 區分하여 裸出實驗한 結果, 작은 잎보다 큰 잎에서 收率이 높은 傾向이었다. 酵素處理 後 裸出된 原形質體의 정제에는 0.8M의 Sucrose가 가장 適當하였다.

• Journal of the Korean Society of Tobacco Science
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• v.8 no.2
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• pp.51-66
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• 1986

Volatile aroma components, non-volatile organic acids in lamina and stems of flue-cured(NC 2326) and burley ( Burley 21) were analyzed by gas chromatography and mass spectrometry, respectively. Then compositional differences of these components between lamina and stems were discussed. The contents of volatile components were higher in flue-cured than in burley tobacco, and it was also higher in lamina then in stem. The major aroma components in lamina were neophytadiene , nicotine, solanone and benzyl alcohol but those in stems were palmitic acid, neophytadiene, nicotine, solanone and phenyl ethyl acetate. On the other hand, the contents of non-volatile organic acids were higher in burley than in flue-cured tobacco, and these values of burley tobacco were higher in lamina than in stem but flue-cured tobacco were higher in stem than in lamina. The major acids in all the above four tabacco samples were malic, citric, oxalic and linolenic acid.

• Journal of Plant Biology
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• v.36 no.1
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• pp.97-104
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• 1993

The lipid composition of thylakoid membranes was compared between mesophyll and bundle sheath chloroplasts of maize. According to mild-denaturing gel electrophoresis, mesophyll thylakoids contained both PS I complex and PS II light-harvesting chlorophyll-protein complex(LHCP), while those of bundle sheath cells contained mainly PS I complex. The amount of lipids per mg chlorophyll was higher in bundle sheath thylakoids than in mesophyll. The major polar lipid classes were monogalactosyldiacylglycerol(MGDG), digalactosyldiacylglycreol, sulfolipid and phosphatidylglycerol (PG) in both tissues. Linolenic acid(18 : 3), linoleic acid(18 : 2) and palmitic acid(16 : 0) were the main fatty acyl components, with higher ratio of unsaturated to saturated fatty acids in bundle sheath thylakoids, suggesting these membranes are more fluid. The most striking difference in lipid composition between the two kinds of tissues was the practical absence of trans- 3-hexadecenoic acid(16 : 1t) in PG of bundle sheath thylakoids. This fatty acid is known to be involved in the association of LHCP as oligomeric form. More than 80% of MGDG molecular species was 18 : 3, 18 : 3, demonstrating that maize is a typical 18 : 3 plant. Therefore, the possibility of the functional relationships between the lamella structure, and thus the distribution of photosystems, and MGDG molecular species was excluded.

• Journal of Plant Biology
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• v.36 no.1
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• pp.105-112
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• 1993

Selection of glyphosate-resistant clones from MNNG-treated mesophyll protoplasts of haploid tobacco and their differentiation were studied. The protoplasts were treated with 0.1 to 100 $\mu\textrm{g}$/mL N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) for 30 min when they expanded to oval shapes. After the treatment, the protoplasts in 4-16 cell stages were transferred to the selective medium containing 1 mM glyphosate for the selection of the glyphosate-resistant colonies. The efficiency of the cell division of the protoplasts in the selective medium decreased as the MNNG concentrations in creased. Optimal MNNG concentration for induction of the glyphosate-resistant clones was 10$\mu\textrm{g}$/mL and mutation frequency was 2.66$\times$10-6. The stability of the glypohsate-resistance of the clones was examined by prolonged subculture in the medium with 1 mM glyphosate, and the resistant clones were survived more than 10 months. Among them one clone has been proliferating and greening and the others were proliferating without greening or greening with slower proliferating.

• Horticultural Science & Technology
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• v.33 no.6
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• pp.820-828
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• 2015

This study was carried out to determine the effects of vernalization, temperature, and soil water deficit (SD) on mesophyll cells, growth, and yield of Chinese cabbage (Brassica campestris L). The palisade parenchyma and spongy tissues of Chinese cabbage were observed under full irrigation and two weeks of SD treatment. These cells were severely collapsed by four weeks SD treatment. The SD treatment had the greatest influence on the growth of Chinese cabbage among the tested treatment factors (vernalization, temperature, and SD), growth significantly decreased by severe drought treatment (four weeks SD treatment). In addition, the relative growth rate, unit leaf rate, leaf area ratio, specific leaf area, and leaf weight ratio were significantly affected by SD treatment; however, other individual factors and their combined treatments did not influence the analyzed growth parameters. The yield under vernalization after high temperature and full irrigation treatments was 3,056 kg/10 a, which was the greatest among all the tested treatments, while four-week SD treatment significantly reduced the yield. Head formation of Chinese cabbage was not altered under SD treatment, and vernalization treatments did not induce bolting. Our results indicated that collapsing mesophyll cells and reduced growth and yield were induced by SD treatment. Thus we suggest that optimal irrigation system should be install to avoid or overcome crippling drought conditions in the open field.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48 no.6
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• pp.506-515
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• 2003

Six barley varieties that showed different degree of drought tolerance were grown with and without drought stress treatment (control), and investigated for the temporal changes in growth and several physiological traits after drought treatment. Soil water potential was -0.05 ㎫ at the initial stage of drought treatment and dropped to -0.29 ㎫ at 19 days after withholding irrigation. Soil water potential (SWP) maintained at -0.05 ㎫ in the control. The dry weight (DW) under the drought treatment were reduced compared to the control as follows: Dicktoo-S (short awn), 69% ; Dicktoo-L (long awn), 70%; Dicktoo-T (tetra), 86%; Dongbori-1, 69%; Suwonssalbori-365, 55% and Tapgolbori, ,37%. Dicktoo lines and Dongbori-1 were more tolerant than Suwonssalbori-365 and Tapgolbori. Leaf relative water contents (RWC) and leaf water potential (LWP) decreased obviously under the drought condition, the decrease being greater especially in the less drought-tolerant barley genotypes. Dongbori-1 and Dicktoo-L in drought treatment showed net photosynthesis of 38% and 17% compared to the control, respectively, and the other four genotypes much lower photosynthesis of 1.1% to 7.0%. Stomatal conductance, mesophyll conductance, and the photochemical efficiency (Fv/Fm) of PS II were reduced by drought treatment, the reduction being greater in drought-sensitive genotypes. The drought-tolerant genotypes had greater osmotic adjustment (OA) capacity under water stress. Thus, the decrease of RWC and LWP was lower and the turgor pressure conservation capacity was higher under water stress in drought-tolerant genotypes. Drought-tolerant genotypes showed less decrease of photosynthesis because stomatal conductance, mesophyll conductance and the ratio (Fv/Fm) of the variable to maximal fluorescence of drought-resistant genotype was decreased less in the drought stress condition. In conclusion, the drought-tolerant genotypes had better water conservation capacity through efficient OA, and this led to the lower decrease of photosynthesis and growth in water stress condition.